Field of the Invention
The present invention relates to the fields of molecular biology and medicine. More specifically, in exemplary embodiments the invention relates to fully engineered nucleic acid superstructures that are engineered to bind and deliver bioactive molecules to target cells or target bioactive molecules to effect medical or clinical treatment.
Description of Related Art
Nucleic acid scaffolds having defined structures are known in the art. For example, U.S. Pat. No. 5,561,043 to Cantor et al. discloses self-assembling nucleic acid aggregates that comprise (1) a first construct comprising a plurality of first single-stranded nucleic acids bound to a first coupling agent forming a first multimer; (2) a second construct comprising a plurality of second single-stranded nucleic acids bound to a second coupling agent forming a second multimer, at least one of the second single-stranded nucleic acids hybridizing with a complementary sequence of one of the first single-stranded nucleic acids; and (3) a plurality of third single-stranded nucleic acids, each of the third single-stranded nucleic acids being attached to a functional group, and each of the third single-stranded nucleic acids hybridizing with a complementary sequence of a single-stranded nucleic acid attached to the aggregate, wherein the first coupling agent and the second coupling agent each include a protein, a 5′-amino-containing oligonucleotide, a 5′-thiol-containing oligonucleotide, a polyamidoamine, or a divalent linker.
U.S. Pat. No. 6,255,469 to Seeman et al. discloses a periodic polynucleic acid structure comprising a lattice of adjacent coplanar repeating units, each repeating unit comprising at least two antiparallel nucleic acid multi-crossover molecules, each of the at least two antiparallel nucleic acid multi-crossover molecules comprising two or more adjacent double helical domains, at least two of the two or more adjacent double helical domains having a first and second cohesive ends, each of the adjacent double helical domains having helix axes in parallel and being connected to adjacent double helical domains at two or more crossover sites, with each antiparallel nucleic acid multi-crossover molecules being connected to an adjacent antiparallel nucleic acid multi-crossover molecule by complementary cohesive ends, thereby forming an extended double helical domain between a first double helical domain of an antiparallel nucleic acid multi-crossover molecule and a second double helical domain of an adjacent antiparallel nucleic acid multi-crossover molecule, wherein a second double helical domain of the antiparallel nucleic acid multi-crossover molecule is not colinear and connectable with a first double helical domain of the adjacent antiparallel nucleic acid crossover molecule to form an extended double helical domain, and wherein each repeating unit is connected to an adjacent repeating unit in the same plane by complementary cohesive ends to form at least one extended double helical domain between adjacent repeating units in the same plane. In addition, methods of making such structures are disclosed. Embodiments of the invention include those in which the structure can include chemically or biologically active molecules.
U.S. Pat. No. 6,814,964 to Virtanen et al. discloses a supramolecule comprising (1) a first supramolecular component having a binding effector molecule covalently joined to at least one first nucleic acid, (2) a second supramolecular component having a therapeutic effector molecule covalently joined to at least one second nucleic acid, wherein at least a portion of the at least one first nucleic acid is hybridized to at least a portion of the at least one second nucleic acid, and wherein the binding effector molecule and the therapeutic effector molecule are selected from the group consisting of proteins, polypeptides, lipids, and sugars. The patent also discloses that the supramolecule can be provided in the form of a pharmaceutical. The patent disclosure is focused on use of such molecules in methods of hydrolyzing viruses.
U.S. Pat. No. 7,223,544 and U.S. Pat. No. 7,799,903 to Luo et al. disclose a method of making a three-dimensional nucleic acid structure formed with a trimer, the method comprising combining a first, a second, and a third polynucleotide in a solution, wherein at least a portion of the first polynucleotide is complementary to at least a portion of the second polynucleotide, wherein at least a portion of the first polynucleotide is complementary to at least a portion of the third polynucleotide; and wherein at least a portion of the second polynucleotide is complementary to at least a portion of the third polynucleotide; and maintaining the solution at conditions effective for the first, second, and third polynucleotides to associate together to form a trimer, wherein each polynucleotide comprises a sticky end, and wherein the sequence of each sticky end is non-complementary. These documents further disclose that the invention described in them provides a solution for a need in the art for branched (Y-shaped) DNA molecules with precisely controlled sizes, which can be incorporated into dendrimer-like DNA (DL-DNA). The DL-DNA is disclosed as useful for, among other things, controlled drug delivery.
U.S. Pat. No. 7,598,363 to Seeman et al. discloses a polynucleic acid structure comprising a polygonal unit whose edges are parallel helices of connected nucleic acid multi-crossover domains along their helix axes, each of the edges having at least one free end with two parallel helices, wherein each of the two parallel helices at one free end of each of the edges terminate in a cohesive end to provide a double cohesive end on the one free end. The patent discloses that the invention described in the document differs from the prior art in that the structures include DNA double crossover (DX) molecules, rather than conventional DNA double helices. Further, the patent discloses that the structures are useful for, among other things, immobilizing enzymes and other catalysts.
U.S. Pat. No. 7,622,567 to Seeman et al. discloses a two dimensional polynucleic acid array of polygonal units linked together by complementary double cohesive ends, comprising a plurality of polygonal units, wherein each of the polygonal units has, as edges, parallel helices of connected nucleic acid multi-crossover domains along their helix axes; each of at least two of the edges of each of the polygonal units has ends with two parallel double helices; each of the two parallel double helices terminate in a cohesive end to provide a double cohesive end at each end of the at least two edges, whereby the double cohesive end of one edge of a polygonal unit is cohered to a complementary double cohesive end of an adjacent polygonal unit in the array to form an extended edge linking together two adjacent polygonal units; and at least one edge, which is different from the at least two edges, of a subset of said polygonal units has at least one end which is attached to a nanoparticle or pendant molecule. The patent discloses that the invention relates to new motifs for polynucleic acid arrays, which is stiffer or more rigid than prior art bulged-junction triangles.
U.S. patent application publication number 2009/0018028 to Lindsay et al., incorporated herein in its entirety by reference, discloses self-assembling, finite nucleic acid tiling arrays, and methods for their synthesis and use. The publication discloses that the invention provides a nucleic acid tiling array comprising a plurality of nucleic acid tiles joined to one another via sticky ends, wherein each nucleic acid tile comprises one or more sticky ends, and wherein a sticky end for a given nucleic acid tile is complementary to a single sticky end of another nucleic acid tile in the nucleic acid tiling array; wherein the nucleic acid tiles are present at predetermined positions within the nucleic acid tiling array as a result of programmed base pairing between the sticky ends of the nucleic acid tiles.
While numerous technologies and nucleic acid nanostructures have been developed to date in the art, none of those nanostructures has been disclosed that has a pre-selected, finite size and that is completely engineered to include specific binding sites for one or more substances that can function in targeting of the nucleic acid nanostructure to a desired target, and delivery of the nanostructure and the attached substance(s) to the target, where target refers to any bioactive molecule, bioactive complex, or biological cell.